The Influence Of Clopidogrel On BMSCs Transplantation For Treatment Of Ischemic Brain Injury

ObjectiveTo evaluate the efficacy of clopidogrel combined with allogeneictransplantation of bone marrow mesenchymal stem cells (BMSCs) compared withBMSCs transplantation or clopidogrel treatment alone in the treatment of middlecerebral artery ischemia (MCAO) models. We explored whether clopidogrel canpromote the effect of BMSCs transplantation on cerebral ischemia and the possiblemechanisms.MethodsBMSCs were cultured by entire medulla from long bones of maleSprague-Dawley rats (200g body weight). P3-5cells were prepared by PBS to thesuspensions of2x106/ml cell. We established the MCAO models, which have beenwell documented to cause cerebral ischemia in rats. The rats that have arrived at thescores were divided randomly into four groups: model group (group B, n=12),clopidogrel group (group C, n=12), BMCSs transplantation group (group D, n=12)and clopidogrel combined with BMCSs transplantation group (group E, n=12). Therats that were sutured after just made a cut in cervical skin and detached to thepterygopalatine artery were selected as the sham group (A, n=12). Group D and Ewere given1ml BMCSs suspensions by tail vein after24hours when the MCAOmodels were established successfully.Group C and E were given clopidogrel25mg/kg/day, group A received no treatment. The neurological score was estimatedafter24hours and on day14. HE staining and TTC were used to measure the infarctionsize. The expression of Bcl-2and Bax in brain tissues were examined by immunohistochemistry.Results1. Neurological function score show that the group B, C, D and E were3.50±0.67,2.25±0.62,2.00±0.66,1.75±0.62. The difference was significant between groupB and other groups (P <0.05), suggesting that the three types of treatment can improvethe function of neurological. Also the difference was significant among the treatmentgroups, suggesting that the effect of group D is better than C, and the effect of group Eis the best (P <0.05).2. Three types of treatment significantly decreased the volume of cerebralinfraction measured by TTC(P <0.05), and the results were209.64±6.52mm3、138.58±4.51mm3、111.35±4.53mm3、77.13±3.70mm3respectively, among them the group Eis the lowest.3. The results of Bcl-2examined by immunohistochemistry showed that group Ato E were8.42±1.03,14.07±1.25,26.03±2.36,32.74±2.35,43.72±3.14(±), group Aas the control. The expression of group E is the most, followed by group D, group B isthe third.4.The results of Bax examined by immunohistochemistry indicated group A to Ewere7.01±0.35,39.84±2.09,29.92±1.89,21.17±1.71,11.36±1.56. Compared withgroup B, the other groups were significantly decreased the expression of Bax(P <0.05).The difference was significant between group E and C or B(P <0.05).Conclusion1.Clopidogrel combined BMSCs transplantation was more effective thanclopidogrel or BMSCs transplantation alone in the treatment of cerebral ischemia andreperfusion injury.2.Synergistic effect of Clopidogrel combined with BMSCs on cerebral ischemiaand reperfusion injury.3.Clopidogrel may through increase the expression of Bcl-2and decrease Bax toimprove the efficacy of BMSCs transplantation in the treatment of cerebral ischemia reperfusion injury.